Isolation and characterization of microsatellite loci in the flour mite Acarus siro

Thirteen microsatellites with dinucleotide repeats were isolated from genomic DNA in solution by hybridization capture. Eleven were evaluated against Acarus siro using a multiple‐tube approach to assess polymorphism and reliability of the results. The multiple‐tube method demonstrated the presence of null alleles due to low levels of DNA. Seven of the 11 microsatellite loci were polymorphic and six produced reliable amplification products, if results only from individuals that scored at all six loci were used in the analysis. These were evaluated in eight strains of A. siro. Two of the samples showed some genetic divergence using F_ST values.     

Energy budget of the grain mite,Acarus siro (Acari: Acaridae)

Immature individuals ofAcarus siro L. ingest a total of 0.1408 J of food energy, 58.5% of which is consumed by the deutonymph, 27.1% by the protonymph and only 14.1% by the larva. The total ingestion is much greater (2.8169 J±0.0981se). Much energy, 0.7031 J±0.0231se, is used for egg production, which is concentrated in the first 20 days of adult life. Assimilation efficiency during this period varies from 63 to 71%. Most energy ingested between the 20th and 40th days of adult life is used in biomass production, and biomass increases significantly up to th 30th day. Afterwards, old individuals expend a large amount of assimilated energy on respiration due to the increased cost of maintenance.Present address: Biology Dept., LB 8042, Georgia Southern University, Statesboro, GA 30460, U.S.A.     

Effectiveness of the BT mite trap for detecting the storage mite pests, Acarus siro, Lepidoglyphus destructor and Tyrophagus longior

Traps have been used extensively to provide early warning of hidden pest infestations. To date, however, there is only one type of trap on the market in the U.K. for storage mites, namely the BT mite trap, or monitor. Laboratory studies have shown that under the test conditions (20 °C, 65% RH) the BT trap is effective at detecting mites for at least 10 days for all three species tested: Lepidoglyphus destructor, Tyrophagus longior and Acarus siro. Further tests showed that all three species reached a trap at a distance of approximately 80 cm in a 24 h period. In experiments using 100 mites of each species, and regardless of either temperature (15 or 20 °C) or relative humidity (65 or 80% RH), the most abundant species in the traps was T. longior, followed by A. siro then L. destructor. Trap catches were highest at 20 °C and 65% RH. Temperature had a greater effect on mite numbers than humidity. Tests using different densities of each mite species showed that the number of L. destructor found in/on the trap was significantly reduced when either of the other two species was dominant. It would appear that there is an interaction between L. destructor and the other two mite species which affects relative numbers found within the trap.The British Crowns right to retain a non-exclusive, royalty-free licence in and to any copyright is acknowledged.This revised version was published online in May 2005 with a corrected cover date.     

A new bioassay method for the detection of resistance to pesticides in the stored product mite Acarus siro (Acari: Acaridae)

The majority of bioassay methods used for determining the susceptibility of the flour mite Acarus siro L. to pesticides provide data which are difficult to assess. In this new method, the mites are confined on treated, moulded filter papers held in position by two glass sheets to form an escape-proof exposure chamber. The method is simple and uses materials that are likely to be readily available. The dose–response data obtained from laboratory and field strains of A. siro to three organophosphorus-based pesticides are presented. The method provided repeatable results and no significant variation when used by different operators. A discriminating dose for the detection of pirimiphos-methyl resistance is suggested.     

Molecular cloning, expression, sequence analyses of dust mite allergen Der f 6 and its IgE-binding reactivity with mite allergic asthma patients in southeast China

We report the cloning and molecular characterization of a full-length cDNA encoding house dust mite allergen, Der f 6 from D. farinae isolated in China. The full-length Der f 6 cDNA was obtained with 840 nucleotides long. Nucleotide sequencing analyses showed a total of 36 mutations in five Der f 6 cDNA clones, corresponding to 23 incompatible amino acid residues. Recombinant Der f 6 (rDer f 6) protein was successfully expressed in and purified from E. coli BL21. Among 20 asthmatic patients, 45% was positive to rDer f 6 by ELISA. Bioinformatics analyses revealed that the mature Der f 6 was a hydrophobic and extracellular protein with chymotrypsin-like serine protease activity, its secondary structure was composed of alpha helix (7.69%), extended strand (34.62%), random coils (57.69%), and the similarity of Der f 6 to Blo t 6, Sui m 6, Der f 3 and Der f 9 was 64, 65, 35, and 38%, respectively.     

Combined effect of an antifeedant α-amylase inhibitor and a predator Cheyletus malaccensis in controlling the stored-product mite Acarus siro

Abstract.  Acarus siro is the most abundant and frequent mite to infest stored-food products, causing allergies and transmitting mycotoxin producing fungi. The predatory mite Cheyletus malaccensis is a candidate species in the biocontrol programme for this pest. In vitro , the α-amylase inhibitor acarbose is effective against the α-amylase of A. siro but not against that of C. malaccensis . In vivo , the impact of acarbose on a population of A. siro is investigated along with the interaction with the predator. Various densities of adult parthenogenetic females of C. malaccensis are reared on A. siro feeding on either a control diet or a diet containing different acarbose concentrations. The combined action of both factors significantly improved the final biocontrol efficiency with C. malaccensis , compensating for the lower energetic content of the prey on acarbose by increasing the number of prey caught. Acarbose had no negative effects on the longevity and the length of oviposition period of C. malaccensis but partially reduced its fecundity. The results are discussed in the context of the integrated control of stored-product mite pests.     

The determination of allergen-specific IgE and IgG antibodies in patients sensitized to the house dust mite Dermatophagoides pteronyssinus

45 patients, hypersensitive to house-dust mites, were examined by the method of skin tests to D. pteronyssinus allergen. Besides, in their blood sera the levels of allergen-specific IgE antibodies were determined in the radioallergosorbent test and allergen-specific IgG antibodies, in the enzyme immunoassay. These tests revealed that in 91% of the patients the results of skin tests were positive, in 68% an elevated level of specific IgE antibodies and in 93% of the patients an elevated level of specific IgG antibodies were detected. All patients showed the positive result in one of the above-mentioned tests. The largest group of the patients (55%) included persons showing the positive result of the skin test and having elevated levels of allergen-specific IgG and IgE antibodies. Thus, in cases of hypersensitivity to house-dust mites the levels of allergen-specific IgG and IgE antibodies in the patients' blood sera should be determined.     

Phenotypic comparison of allergic airway responses to house dust mite in three rat strains

Brown Norway (BN) rats develop a robust response to antigens in the lung, characterized by a large increase in allergen-specific immune function and pulmonary eosinophilia. The objective of this study was to investigate alternative models by determining whether other rat strains could be sensitized to house dust mite (HDM) antigen and whether the allergic disease process could be worsened with repeated allergen exposure. In general, BN rats sensitized by either subcutaneous or intratracheal routes exhibited increased pulmonary allergy compared with Sprague-Dawley (SD) and Lewis (L) rats. Multiple intratracheal allergen exposures incrementally increased HDM-specific immune function in BN rats but progressively decreased eosinophil recruitment and markers of lung injury. SD rats had more moderate responses, whereas L rats were relatively unresponsive. Because BN rats developed stronger clinical hallmarks of allergic asthma under various immunization regimes compared with SD and L rats, we conclude that the BN is the most appropriate strain for studying allergic asthma-like responses in rats. Phenotypic differences in response to HDM were associated with differences in the Th1/Th2 cytokine balance and antioxidant capacity.     

Ultrastructure of the digestive tract in Acarus siro (Acari: Acaridida)

The gut of the mite Acarus siro is characterized on the ultrastructural level. It consists of the foregut (pharynx, esophagus), midgut (ventriculus, caeca, colon, intercolon, postcolonic diverticula, postcolon), and hindgut (anal atrium). The gut wall is formed by a single-layered epithelium; only regenerative cells are located basally and these have no contact with the lumen. Eight cell types form the whole gut: (i) simple epithelial cells forming fore- and hindgut; (ii) cells that probably produce the peritrophic membrane; (iii) regenerative cells occurring in the ventriculus, caeca, colon, and intercolon; (iv) spherite cells and (v) digestive cells forming the ventriculus and caeca; (vi) colonic cells and (vii) intercolonic cells; and (viii) cells forming the walls of postcolonic diverticula and postcolon. Spherite and digestive cells change in structure during secretory cycles, which are described and discussed. The cycle of spherite, colonic, and intercolonic cells is terminated by apoptosis. Ingested food is packed into a food bolus surrounded by a single homogeneous peritrophic membrane formed by addition of lamellae that subsequently fuse together. The postcolonic diverticula serve as a shelter for filamentous bacteria, which also are abundant in the intercolon.     

Epigenetic changes in B lymphocytes associated with house dust mite allergic asthma

Although there is no doubt about the influence of the genetic background in the onset of the allergic diseases, Epigenome-Wide Association Studies are needed to elucidate the possible relationship between allergic diseases and epigenomic dysregulation. In this study we aimed to analyze the epigenetic patterns, in terms of DNA methylation, of three well-characterized populations of house dust mite allergic subjects, aspirin-intolerant asthmatics and controls. As a first, genome-wide phase, we used the HELP assay to study the methylation patterns in CD19⁺ B lymphocytes in these populations, and found that there are reproducible epigenetic differences at limited numbers of loci distinguishing the groups, corroborated by bisulphite MassArray in a second validation phase of an expanded 40 subject group. These validated epigenetic changes occur at loci characterized as important for the immune response. One such locus is a new candidate gene, CYP26A1, which shows differential methylation patterns and expression levels between groups. Our results suggest that epigenomic dysregulation may contribute to the susceptibility to allergic diseases, showing for the first time differences in DNA methylation between allergic and non-allergic healthy subjects, both globally and at specific loci. These observations indicate that the epigenome may offer new pathophysiological insights and therapeutic targets in atopic diseases.     

Epigenetic changes in B lymphocytes associated with house dust mite allergic asthma

Although there is no doubt about the influence of the genetic background in the onset of the allergic diseases, Epigenome-Wide Association Studies are needed to elucidate the possible relationship between allergic diseases and epigenomic dysregulation. In this study we aimed to analyze the epigenetic patterns, in terms of DNA methylation, of three well-characterized populations of house dust mite allergic subjects, aspirin-intolerant asthmatics and controls. As a first, genome-wide phase, we used the HELP assay to study the methylation patterns in CD19 (+) B lymphocytes in these populations, and found that there are reproducible epigenetic differences at limited numbers of loci distinguishing the groups, corroborated by bisulphite MassArray in a second validation phase of an expanded 40 subject group. These validated epigenetic changes occur at loci characterized as important for the immune response. One such locus is a new candidate gene, CYP26A1, which shows differential methylation patterns and expression levels between groups. Our results suggest that epigenomic dysregulation may contribute to the susceptibility to allergic diseases, showing for the first time differences in DNA methylation between allergic and non-allergic healthy subjects, both globally and at specific loci. These observations indicate that the epigenome may offer new pathophysiological insights and therapeutic targets in atopic diseases.     

Effects of radiation on spermatogenesis in Acarus siro L. (Acari: Acaridae).

Modifications of spermatogenesis and anatomy of male reproductive tracts caused by radiation (fast electrons) are described in Acarus siro and compared with data from other arthropod taxa. Radiation induced severe changes in gonial cells, including dilatation of endoplasmic reticulum and disruption of mitochondrial cristae. The latter sometimes created roundish myelin structures inside the mitochondrial matrix. Damage in developing cells caused an interruption of spermatogenesis and reduced the number of spermatids produced; high doses can cause sterility.     

Pirimiphos-methyl resistance in two stored product mites, Acarus siro and Acarus farris, as detected by impregnated paper bioassay and esterase activity assays

The response to pirimiphos-methyl, in one strain of Acarus farris and two strains of Acarus siro, was assessed using an impregnated filter paper bioassay and by the selection of adults following exposure to pirimiphos-methyl. It was concluded that one of the strains of A. siro was resistant to pirimiphos-methyl and that a major resistance mechanism was involved. The second strain of A. siro gave a response similar to that of a laboratory strain unexposed to organophosphates and was considered to be susceptible. The A. farris strain responded to selection at the ED50 but not at the ED99, and it was concluded that a minor resistance mechanism is present in this strain. Assays of esterase activity were used to attempt to identify the biochemical mechanisms involved in the resistance detected by the bioassays. The A. farris and susceptible A. siro strains showed similar levels of esterase activity but the esterase activity of the resistant A. siro strain was significantly greater. An increase in esterase activity followed selection of both the A. farris strain and the resistant A. siro strain. An acetylcholinesterase assay showed no significant difference between the susceptible and pirimiphos-methyl selected strains of A. siro. The results suggest that esterases are involved in the resistance to pirimiphos-methyl found in A. siro and A. farris but that in A. siro, at least, other mechanisms may also be present.     

Induction of allergic reactions in guinea pigs with purified house dust mite allergens

To establish a guinea pig model for house dust mite allergy with purified mite allergens, we studied the immune response to two major mite allergens, native Der f 1 (nDer f 1) and recombinant Der f 2 (rDer f 2) and crude mite extract in Hartley guinea pigs. Animals were immunized with either mite extract, nDer f 1 or rDer f 2, four times at 2- to 3-week intervals. Then the guinea pigs were examined as to the status of sensitization to the sensitizing antigen. Intradermal injection of mite antigens to mite extract-, nDer f 1-, and rDer f 2-sensitized animals induced both immediate and late-phase cutaneous reactions. Allergic airway disease was also provoked by the intranasal instillation of rDer f 2 or mite extract. Anti-nDer f 1 and -rDer f 2 IgE as well as anti-mite extract IgE were produced in the sensitized guinea pigs and IgE titer for three mite antigens were comparable. We concluded that immunization of Hartley guinea pigs with nDer f 1 and rDer f 2 achieved sensitization to mite allergens, which was comparable to that obtained by the immunization with mite extract. A mite-allergic model suitable for immunological and pharmacological studies was established from rDer f 2-sensitized guinea pigs.     

Comparison of detection methods for Acarus siro (Acari: Acaridida: Acarididae) contamination in grain

Acarus siro L. 1758 (Acari: Acaridida: Acarididae) is an important pest of stored grain because it contaminates the grain by allergens and transfers pathogenous microorganisms. Rapid detection of contamination enables to intercept an early grain infestation by the pest. In this study, we compared the usability and efficiency of various detection approaches. Under laboratory conditions, grain samples of various sizes were infested by different levels of the following contaminants: eggs, adults, and feces of A. siro. The samples were analyzed by enzyme-linked immunosorbent assay (ELISA) by using anti-A. siro polyclonal antibody (immunochemical method), extracted in Berlese-Tullgren funnels, sieved, and processed by filth-flotation (conventional methods). The adults or juveniles of A. siro could be detected by all the three tested conventional methods and ELISA with detection limits in the range from 221 to 1,157 mites/kg grain. Eggs were detected by filth-flotation only; the detection limit was 1,950 eggs/kg grain. The feces of A. siro were detectable by ELISA test, only. ELISA enabled the detection of the feces with the minimal threshold level of 1.04 microg feces/g grain; it means the assay allowed to trace less than one metabolically active mite per gram of grain. The study thus demonstrated that reliable A. siro detection in grain can only be achieved by combining different detection methods. European Union and U.S. administratives dictate zero or near-zero tolerance level for mite infestation in stored products. This demand is difficult to fulfill, because every detection method is limited by its detection limit; thus, it is hard to reliably detect infestation levels lower than obtained detection limits. This methodical limitation is discussed in context with the determined detection limits of the tested methods.     

Prevention of house dust mite induced allergic airways disease in mice through immune tolerance

Allergic airways disease is a consequence of a Th2 response to an allergen leading to a series of manifestations such as production of allergen-specific IgE, inflammatory infiltrates in the airways, and airway hyper-reactivity (AHR). Several strategies have been reported for tolerance induction to allergens leading to protection from allergic airways disease. We now show that CD4 blockade at the time of house dust mite sensitization induces antigen-specific tolerance in mice. Tolerance induction is robust enough to be effective in pre-sensitized animals, even in those where AHR was pre-established. Tolerant mice are protected from airways eosinophilia, Th2 lung infiltration, and AHR. Furthermore, anti-CD4 treated mice remain immune competent to mount immune responses, including Th2, to unrelated antigens. Our findings, therefore, describe a strategy for tolerance induction potentially applicable to other immunogenic proteins besides allergens.     

Intranasal exposure of mice to house dust mite elicits allergic airway inflammation via a GM-CSF-mediated mechanism

It is now well established that passive exposure to inhaled OVA leads to a state of immunological tolerance. Therefore, to elicit allergic sensitization, researchers have been compelled to devise alternative strategies, such as the systemic delivery of OVA in the context of powerful adjuvants, which are alien to the way humans are exposed and sensitized to allergens. The objectives of these studies were to investigate immune-inflammatory responses to intranasal delivery of a purified house dust mite (HDM) extract and to evaluate the role of GM-CSF in this process. HDM was delivered to BALB/c mice daily for 10 days. After the last exposure, mice were killed, bronchoalveolar lavage was performed, and samples were obtained. Expression/production of Th2-associated molecules in the lymph nodes, lung, and spleen were evaluated by real-time quantitative PCR and ELISA, respectively. Using this exposure protocol, exposure to HDM alone generated Th2 sensitization based on the expression/production of Th2 effector molecules and airway eosinophilic inflammation. Flow cytometric analysis demonstrated expansion and activation of APCs in the lung and an influx of activated Th2 effector cells. Moreover, this inflammation was accompanied by airways hyper-responsiveness and a robust memory-driven immune response. Finally, administration of anti-GM-CSF-neutralizing Abs markedly reduced immune-inflammatory responses in both lung and spleen. Thus, intranasal delivery of HDM results in Th2 sensitization and airway eosinophilic inflammation that appear to be mediated, at least in part, by endogenous GM-CSF production.     

Sensitivity to house dust mite allergens and prevalence of allergy-causing house dust mite species in Pothwar,Pakistan

This study is the first report on the epidemiological status of house dust mite (HDM) allergy in Pothwar, Pakistan. Allergy data of 2087 symptomatic patients were obtained, of whom 1706 (81.7%) patients were skin-prick-test positive for HDM allergens. This percentage was significantly higher than for pollen and food allergens. In the results of this study Dermatophagoides farinae (61%) and D. pteronyssinus (29%) were the predominant species in the study area. Besides these pyroglyphids, predatory Cheyletus sp. (10%) and an oribatid mite sp. (1%) were also observed. Random and patients’ houses showed 87.4 and 87.1% positive mite infestation, respectively. Mean (±?SEM) D. farinae counts per g of dust in random samples was 235.4 ± 7.93 compared to 274.7 ± 10.78 from patients’ homes. Mean D. pteronyssinus counts from random houses compared to patients’ houses were 115.0 ± 4.57 and 124.6 ± 5.76, respectively. Mite counts depicted seasonal variation, with peaks during monsoon season. ELISA results of dust samples demonstrated that of the dust samples with?>?10 µg/g of dust, the threshold value described as a risk factor for developing asthma, 57.6% had Der f1 and 20% Der p1 allergen load. Mean Der f1 burden was significantly higher than Der p1, with maximum levels during monsoon and autumn seasons. This research established a better awareness about the epidemiological status of HDM allergy and prevalence of allergy causing HDM species in Pakistan.